Project information
Rational re-design of microbial enzymes involved in degradation of toxic organic pollutants.
- Project Identification
- ME 276
- Project Period
- 6/1998 - 12/2002
- Investor / Pogramme / Project type
-
Ministry of Education, Youth and Sports of the CR
- Programme KONTAKT (ME+MEB)
- MU Faculty or unit
- Faculty of Science
- Keywords
- protein engineering, haloalkane dehalogenase, site-directed mutagenesis, biodegradation, detoxification, structure-function relationships, computer modelling
The main objective of the project is the investigation of structure-function relationships of the enzymes involved in degradation of toxic substances - halogenated aliphatic hydrocarbons. The structures of the proteins from the family of haloalkane dehalogenases will be rationaly modified using the methods of genetic engineering. The impact of the changes in proteins structure on activity and specificity will be tested using steady-state kinetic measurements. Design of the mutant proteins will be conducted using computer modelling. Mutant proteins will be constructed using site-directed mutagenesis techniques. General structure-function relationships will be formulated from derived experimental data.
Publications
Total number of publications: 18
2002
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Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene
Applied and Environmental Microbiology, year: 2002, volume: 68, edition: 7
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Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition
Biochemistry, year: 2002, volume: 41, edition: 15
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Halide-stabilizing residues of haloalkane dehalogenases studied by quantum mechanic calculations and site-directed mutagenesis
Biochemistry, year: 2002, volume: 41, edition: 48
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Impact of Orthogonal Signal Correction (OSC) on the predictive ability of CoMFA models for the ciliate toxicity of nitrobenzenes
Quantitative Structure-Activity Relationships, year: 2002, volume: 21, edition: 1
2001
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Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: characterization of the haloalkane dehalogenase mutants
Journal of Microbiological Methods, year: 2001, volume: 44, edition: 1
-
Molecular dissection of interactions between Rad51 and members of the recombination-repair group
Molecular and Cellular Biology, year: 2001, volume: 21, edition: 3
-
Reaction mechanism and stereochemistry of gamma-hexachlorocyclohexane dehydrochlorinase LinA
Journal of Biological Chemistry, year: 2001, volume: 276, edition: 11
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TRITON: graphic software for rational engineering of enzymes
Trends in Biochemical Sciences, year: 2001, volume: 28, edition: 1
2000
-
Dehalogenation of haloalkanes by Mycobacterium tuberculosis H37Rv and other mycobacteria
Applied and Environmental Microbiology, year: 2000, volume: 66, edition: 1
-
Effect of the carbon source on assessment of degrading bacteria with the spread-plating technique during in situ bioremediation
Folia Microbiologica, year: 2000, volume: 45, edition: 1